Most glycosaminoglycan (GAG)-defective mutants have been isolated and characterized from Chinese hamster ovary (CHO) cells. Wild-type and GAG-defective CHO cells have been used by several hundreds of laboratories to study how altering the GAG structure of proteoglycans affects fundamental properties of cells, such as bacterial/viral infection, signaling, protein degradation, and cell adhesion. This chapter describes methods used to construct and characterize new CHO cell lines with gain-of-function GAG structures. These novel CHO cell lines allow herpes simplex virus (HSV) entry or have anticoagulant properties that are not possessed by wild-type CHO cells. The method used to study GAG biosynthetic mechanisms that control specific GAG sequence assembly is also described. © 2006 Elsevier Inc. All rights reserved.
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